DOCSLIB.ORG

LARP6 Part of This Dissertation

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
LARP6 Part of This Dissertation Florida State University Libraries Electronic Theses, Treatises and Dissertations The Graduate School 2013 Characterization of Mechanisms Controlling Translational Regulation of Type I Collagen in Liver Fibrosis as Target for Novel Anti- Fibrotics and the Regulation of Neural Tube Closure during Embryonic Development Zarko Manojlovic Follow this and additional works at the FSU Digital Library. For more information, please contact [email protected] THE FLORIDA STATE UNIVERSITY COLLEGE OF MEDICINE CHARACTERIZATION OF MECHANISMS CONTROLLING TRANSLATIONAL REGULATION OF TYPE I COLLAGEN IN LIVER FIBROSIS AS TARGET FOR NOVEL ANTI-FIBROTICS AND THE REGULATION OF NEURAL TUBE CLOSURE DURING EMBRYONIC DEVELOPMENT By ZARKO MANOJLOVIC A Dissertation submitted to the Department of Biomedical Sciences in partial fulfillment of the requirements for the degree of Doctor of Philosophy Degree Awarded: Summer Semester, 2013 Zarko Manojlovic defended this dissertation on April 22, 2013. The members of the supervisory committee were: Branko Stefanovic Professor Directing Dissertation Zuoxin Wang University Representative Myra Hurt Committee Member Yoichi Kato Committee Member James Olcese Committee Member The Graduate School has verified and approved the above-named committee members, and certifies that the dissertation has been approved in accordance with university requirements. ii I dedicate this work to my loving family that had to endure many struggles for this opportunity of a lifetime. iii ACKNOWLEDGEMENTS In my opinion, the process of scientific discovery is a glorious expansion of our intellect and it is just as important as the discovery itself. This sensational quest for answers is ultimately an endless collaborative effort that leads to an integrated web of informatics. First and foremost, a simple thank you is not enough to express the gratitude and the greatest appreciation that I have for my major professor Dr. Branko Stefanovic whose virtuosity and the principle of a strong foundation allowed me to progress towards a successful scientific career. He is the mastermind behind this exciting project, and a great teacher not just of science, but also of life. With wit, Dr. Stefanovic eviscerates experimental issues leading to a very productive environment. He provided me with tools to become a critical thinker and a skilled scientist. His teachings on proficiency and emphasis on quality will be a skill that I will embrace for years to come. Special thanks to all members of my doctoral committee, Drs. Myra Hurt, James Olcese, Yoichi Kato and Zuoxin Wang for their dedication, guidance and professional advice throughout my graduate career. Many thanks to Lela Stefanovic “mother of the lab” for her brilliant technical support and for keeping the lab functional. It was a great pleasure to be working with an exceptional group of lab members, Hao Wang, Yujie Zhang, Drs. Le Cai, Azariyas Challa and Milica Vukmirovic. I would also like to thank Dr. Richard Nowakowski for his expertise and advise on professional development and leadership, Ruth Didier for all the help with the confocal and FACS, the entire administrational personnel for all the help with non-scientific matters, Lilly Lewis for excellent help with academic support in making not just my life, but also life of every graduate student much easier, Jennifer Sweetman for all the hard work and the endless uphill battle with the university in regards to my NIH-NRSA funds, Dr. Ewa Bienkiewicz for help with proteomics, Dr. John Blackmon for his expertise in histology analysis, Dr. Akash Gunjan for help and advice in ubiquitin studies, Dr. Timothy Megraw for the advice on cilia and confocal aspects of my FISH study, a very special thanks to Oscar Cabrera who is not only a true friend, but also a great colleague that shared his expertise on confocal software including FISH image acquisition. Special thanks to the entire Biomedical Sciences family. iv Furthermore, I would like to thank Dr. Kato and his laboratory members, Akiko Kato and Koichi Tanaka for a great experience working together closely on the LARP6 part of this dissertation. My thanks also to Dr. Chee-Gun Lee for the generous gift of RHA clone, to Dr. Kathleen Boris-Lawrie for generous information in regards to siRNAs (RHA) required for my RNA Helicase A study, the entire ACUC staff for making all my in-vivo experiments possible, Dr. Blaine Bartholomew that helped in shaping my early scientific career and Drs. Bryant Chase and Thomas Keller for their support with my NIH-F31 grant. It is without slightest doubt in my mind that this journey would have never been possible without the help of a former boss and a very special friend, Dr. Carlos Bolaṅos. I will always be grateful for his patience and guidance that opened doors for future advancements. It is in his lab that I was introduced to an excellent and exceptional student, Sergio Iniquez. Their influences and dedication to brilliance helped me flourish into a highly competitive and ambitious researcher. Even though my tenure in his lab was short, an eternal friendship was developed. A standing ovation to my father (Vlatko Manojlovic) and my sweetheart of a mother (Snjezana Manojlovic) for dedicating their lives for me and my brother (Marko Manojlovic). Despite the fact that we endured a past filled with cruelty, embraced with war, destruction, and endless refuge, they kept us together. They chose to come to this great country unselfishly, to give me and my little brother an opportunity for a better life and in the process sacrificed their professional careers, friends and everything they knew. They are all I have left and I love them. A very special thank you to my first teacher Vesna Vukancic, little did she know that I will be a student for life, to my wonderful aunt Jelena Manojlovic who I idolize greatly, Slavica Suljuzovic for her unconditional love and support and deceased Dr. Stambak, who I admired greatly and as a result initiated my scientific journey at age of 6. I do not know how this process would have been possible without my wonderful girlfriend, my loving future wife and most importantly my best friend, Heather Case. She is my better half and an escape from science. She is everything I could ever ask for, plus more. Thank you for keeping me sane and for loving me. This work was supported by the R01 NIH [5R01DK059466-08] to B.S, and F31 NIH/NIAAA NRSA [AA019845-01A1] to Z.M. v TABLE OF CONTENTS List of Tables ............................................................................................................................... viii List of Figures ................................................................................................................................ ix Abstract .......................................................................................................................................... xi 1. INTRODUCTION ...................................................................................................................1 1.1 - Abbreviations.........................................................................................................1 1.2 - Pathological Process of Liver Fibrosis ..................................................................1 1.3 - Molecular Mechanism of Type I Collagen Expression .........................................3 1.4 - Anti-fibrotic Treatment Approaches and Future Directions................................11 1.5 – Role of LARP6 in embryonic development........................................................12 2. ROLE OF RNA HELICASE A IN TRANSLATION REGULATION OF TYPE I COLLAGEN MRNAS ...................................................................................................................14 2.1 - Introduction .........................................................................................................14 2.2 - Results .................................................................................................................16 2.3 - Discussion ............................................................................................................31 2.4 - Materials and Methods ........................................................................................36 3. A CRITICAL ROLE OF IMMUNOPHILIN FKBP3 IN TYPE I COLLAGEN SYNTHESIS ..................................................................................................................................42 3.1 - Introduction .........................................................................................................42 3.2 - Results .................................................................................................................43 3.3 - Discussion ............................................................................................................57 3.4 - Materials and Methods ........................................................................................60 4. FK506 PREVENTS EARLY STAGES OF ETHANOL INDUCED HEPATIC FIBROSIS BY TARGETING LARP6 DEPENDENT MECHANISM OF COLLAGEN SYNTHESIS ........65 4.1 - Introduction .........................................................................................................65 4.2 - Results .................................................................................................................68 4.3 - Discussion ............................................................................................................83 4.4 - Materials and Methods ........................................................................................87
Load more

Recommended publications
  • Prostate Cancer Prognostics Using Biomarkers Prostatakrebsprognostik Mittels Biomarkern Prognostic Du Cancer De La Prostate Au Moyen De Biomarqueurs
    (19) TZZ Z_T (11) EP 2 885 640 B1 (12) EUROPEAN PATENT SPECIFICATION (45) Date of publication and mention (51) Int Cl.: of the grant of the patent: G01N 33/574 (2006.01) C12Q 1/68 (2018.01) 18.07.2018 Bulletin 2018/29 C40B 30/04 (2006.01) (21) Application number: 13829137.2 (86) International application number: PCT/US2013/055429 (22) Date of filing: 16.08.2013 (87) International publication number: WO 2014/028884 (20.02.2014 Gazette 2014/08) (54) PROSTATE CANCER PROGNOSTICS USING BIOMARKERS PROSTATAKREBSPROGNOSTIK MITTELS BIOMARKERN PROGNOSTIC DU CANCER DE LA PROSTATE AU MOYEN DE BIOMARQUEURS (84) Designated Contracting States: • GHADESSI, Mercedeh AL AT BE BG CH CY CZ DE DK EE ES FI FR GB New Westminster, British Columbia V3M 6E2 (CA) GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO • JENKINS, Robert, B. PL PT RO RS SE SI SK SM TR Rochester, Minnesota 55902 (US) • VERGARA CORREA, Ismael A. (30) Priority: 16.08.2012 US 201261684066 P Bundoora, Victoria 3083 (AU) 13.02.2013 US 201361764365 P 14.03.2013 US 201361783124 P (74) Representative: Cornish, Kristina Victoria Joy et al Kilburn & Strode LLP (43) Date of publication of application: Lacon London 24.06.2015 Bulletin 2015/26 84 Theobalds Road London WC1X 8NL (GB) (73) Proprietors: • Genomedx Biosciences, Inc. (56) References cited: Vancouver BC V6B 2W9 (CA) WO-A1-2009/143603 WO-A1-2013/090620 • MAYO FOUNDATION FOR MEDICAL WO-A2-2006/091776 WO-A2-2006/110264 EDUCATION AND RESEARCH WO-A2-2007/056049 US-A1- 2006 134 663 Rochester, MN 55905 (US) US-A1- 2007 037 165 US-A1- 2007 065 827 US-A1-
    [Show full text]
  • Naringenin Regulates FKBP4/NR3C1/TMEM173 Signaling Pathway in Autophagy and Proliferation of Breast Cancer and Tumor-Infltrating Dendritic Cell Maturation
    Naringenin Regulates FKBP4/NR3C1/TMEM173 Signaling Pathway in Autophagy and Proliferation of Breast Cancer and Tumor-Inltrating Dendritic Cell Maturation Hanchu Xiong ( [email protected] ) Zhejiang Provincial People's Hospital https://orcid.org/0000-0001-6075-6895 Zihan Chen First Hospital of Zhejiang Province: Zhejiang University School of Medicine First Aliated Hospital Baihua Lin Zhejiang Provincial People's Hospital Cong Chen Zhejiang University School of Medicine Sir Run Run Shaw Hospital Zhaoqing Li Zhejiang University School of Medicine Sir Run Run Shaw Hospital Yongshi Jia Zhejiang Provincial People's Hospital Linbo Wang Zhejiang University School of Medicine Sir Run Run Shaw Hospital Jichun Zhou Zhejiang University School of Medicine Sir Run Run Shaw Hospital Research Keywords: FKBP4, TMEM173, Autophagy, Exosome, Dendritic cell, Breast cancer Posted Date: July 7th, 2021 DOI: https://doi.org/10.21203/rs.3.rs-659646/v1 License: This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Page 1/38 Abstract Background TMEM173 is a pattern recognition receptor detecting cytoplasmic nucleic acids and transmits cGAS related signals that activate host innate immune responses. It has also been found to be involved in tumor immunity and tumorigenesis. Methods Bc-GenExMiner, PROMO and STRING database were used for analyzing clinical features and interplays of FKBP4, TMEM173 and NR3C1. Transient transfection, western blotting, quantitative real-time PCR, luciferase reporter assay, immunouorescence and nuclear and cytoplasmic fractionation were used for regulation of FKBP4, TMEM173 and NR3C1. Both knockdown and overexpression of FKBP4, TMEM173 and NR3C1 were used to analyze effects on autophagy and proliferation of breast cancer (BC) cells.
    [Show full text]
  • De Novo Phosphatidylcholine Synthesis in Intestinal Lipid Metabolism and Disease
    De Novo Phosphatidylcholine Synthesis in Intestinal Lipid Metabolism and Disease by John Paul Kennelly A thesis submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Nutrition and Metabolism Department of Agricultural, Food and Nutritional Science University of Alberta © John Paul Kennelly, 2018 Abstract Phosphatidylcholine (PC), the most abundant phospholipid in eukaryotic cells, is an important component of cellular membranes and lipoprotein particles. The enzyme CTP: phosphocholine cytidylyltransferase (CT) regulates de novo PC synthesis in response to changes in membrane lipid composition in all nucleated mammalian cells. The aim of this thesis was to determine the role that CTα plays in metabolic function and immune function in the murine intestinal epithelium. Mice with intestinal epithelial cell-specific deletion of CTα (CTαIKO mice) were generated. When fed a chow diet, CTαIKO mice showed normal lipid absorption after an oil gavage despite a ~30% decrease in small intestinal PC concentrations relative to control mice. These data suggest that biliary PC can fully support chylomicron output under these conditions. However, when acutely fed a high-fat diet, CTαIKO mice showed impaired intestinal fatty acid and cholesterol uptake from the intestinal lumen into enterocytes, resulting in lower postprandial plasma triglyceride concentrations. Impaired intestinal fatty acid uptake in CTαIKO mice was linked to disruption of intestinal membrane lipid transporters (Cd36, Slc27a4 and Npc1l1) and higher postprandial plasma Glucagon-like Peptide 1 and Peptide YY. Unexpectedly, there was a shift in expression of bile acid transporters to the proximal small intestine of CTαIKO mice, which was associated with enhanced biliary bile acid, PC and cholesterol output relative to control mice.
    [Show full text]
  • Taylor Et Al REVISED MS28926-1.Pdf
    1 The effect of micronutrient supplementation on growth and hepatic 2 metabolism in diploid and triploid Atlantic salmon (Salmo salar) parr 3 fed a low marine ingredient diet 4 5 John F. Taylor a*, Luisa M. Vera a, Christian De Santis a, Erik-Jan Lock b, Marit Espe b, 6 Kaja H. Skjærven b, Daniel Leeming c, Jorge del Pozo d, Jose Mota-Velasco e, Herve 7 Migaud a, Kristin Hamre b, Douglas R. Tocher a 8 9 a Institute of Aquaculture, University of Stirling, Stirling, FK9 4LA, UK 10 b Institute of Marine Research, PO box 1870 Nordnes, 5817 Bergen, Norway 11 c BioMar Ltd., Grangemouth, FK3 8UL, UK 12 d The Royal (Dick) School of Veterinary Studies, Edinburgh, EH25 9RG, UK 13 e Hendrix Genetics, Landcatch Natural Selection Ltd., Lochgilphead, PA31 8PE, UK 14 15 Running Title: Dietary micronutrient supplementation in Atlantic salmon 16 17 ms. has 31 pg.s, 4 figures, 9 tables, 4 suppl. files 18 19 Corresponding Author: 20 Dr John F. Taylor 21 Institute of Aquaculture, University of Stirling, Stirling, FK9 4LA, UK 22 Tel: +44-01786 467929 ; Fax: +44-01768 472133 23 [email protected] 24 Accepted refereed manuscript of: Taylor JF, Vera LM, De Santis C, Lock E, Espe M, Skjaerven KH, Leeming D, Del Pozo J, Mota-Velasco J, Migaud H, Hamre K & Tocher DR (2019) The effect of micronutrient supplementation on growth and hepatic metabolism in diploid and triploid Atlantic salmon (Salmo salar) parr fed a low marine ingredient diet. Comparative Biochemistry and Physiology. Part B, Biochemistry and Molecular Biology, 227, pp.
    [Show full text]
  • Supplementary Table S4. FGA Co-Expressed Gene List in LUAD
    Supplementary Table S4. FGA co-expressed gene list in LUAD tumors Symbol R Locus Description FGG 0.919 4q28 fibrinogen gamma chain FGL1 0.635 8p22 fibrinogen-like 1 SLC7A2 0.536 8p22 solute carrier family 7 (cationic amino acid transporter, y+ system), member 2 DUSP4 0.521 8p12-p11 dual specificity phosphatase 4 HAL 0.51 12q22-q24.1histidine ammonia-lyase PDE4D 0.499 5q12 phosphodiesterase 4D, cAMP-specific FURIN 0.497 15q26.1 furin (paired basic amino acid cleaving enzyme) CPS1 0.49 2q35 carbamoyl-phosphate synthase 1, mitochondrial TESC 0.478 12q24.22 tescalcin INHA 0.465 2q35 inhibin, alpha S100P 0.461 4p16 S100 calcium binding protein P VPS37A 0.447 8p22 vacuolar protein sorting 37 homolog A (S. cerevisiae) SLC16A14 0.447 2q36.3 solute carrier family 16, member 14 PPARGC1A 0.443 4p15.1 peroxisome proliferator-activated receptor gamma, coactivator 1 alpha SIK1 0.435 21q22.3 salt-inducible kinase 1 IRS2 0.434 13q34 insulin receptor substrate 2 RND1 0.433 12q12 Rho family GTPase 1 HGD 0.433 3q13.33 homogentisate 1,2-dioxygenase PTP4A1 0.432 6q12 protein tyrosine phosphatase type IVA, member 1 C8orf4 0.428 8p11.2 chromosome 8 open reading frame 4 DDC 0.427 7p12.2 dopa decarboxylase (aromatic L-amino acid decarboxylase) TACC2 0.427 10q26 transforming, acidic coiled-coil containing protein 2 MUC13 0.422 3q21.2 mucin 13, cell surface associated C5 0.412 9q33-q34 complement component 5 NR4A2 0.412 2q22-q23 nuclear receptor subfamily 4, group A, member 2 EYS 0.411 6q12 eyes shut homolog (Drosophila) GPX2 0.406 14q24.1 glutathione peroxidase
    [Show full text]
  • FKBP4 Is a Malignant Indicator in Luminal a Subtype of Breast Cancer
    Journal of Cancer 2020, Vol. 11 1727 Ivyspring International Publisher Journal of Cancer 2020; 11(7): 1727-1736. doi: 10.7150/jca.40982 Research Paper FKBP4 is a malignant indicator in luminal A subtype of breast cancer Hanchu Xiong1,2*, Zihan Chen3*, Wenwen Zheng2, Jing Sun2, Qingshuang Fu4, Rongyue Teng1, Jida Chen1, Shuduo Xie1, Linbo Wang1, Xiao-Fang Yu2, Jichun Zhou1 1. Department of Surgical Oncology, Sir Run Run Shaw Hospital, Zhejiang University, Hangzhou, 310016, China. 2. Cancer Institute, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, 310016, China. 3. Surgical Intensive Care Unit, First Affiliated Hospital, Zhejiang University, Hangzhou, Zhejiang, 310016, China. 4. Rui An Hospital of Traditional Chinese Medicine, Wenzhou, 325200, China. * These authors contributed equally Corresponding authors: Xiao-Fang Yu, E-mail: [email protected]; Jichun Zhou, E-mail: [email protected] © The author(s). This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/). See http://ivyspring.com/terms for full terms and conditions. Received: 2019.10.08; Accepted: 2019.12.20; Published: 2020.01.16 Abstract Purpose: FKBP4 is a member of the immunophilin protein family, which plays a role in immunoregulation and basic cellular processes involving protein folding and trafficking associated with HSP90. However, the relationship between abnormal expression of FKBP4 and clinical outcome in luminal A subtype breast cancer (LABC) patients remains to be elucidated. Methods: Oncomine, bc-GenExMiner and HPA database were used for data mining and analyzing FKBP4 and its co-expressed genes. GEPIA database was used for screening co-expressed genes of FKBP4.
    [Show full text]
  • FKBP3 Rabbit Pab
    Leader in Biomolecular Solutions for Life Science FKBP3 Rabbit pAb Catalog No.: A6907 Basic Information Background Catalog No. The protein encoded by this gene is a member of the immunophilin protein family, A6907 which play a role in immunoregulation and basic cellular processes involving protein folding and trafficking. This encoded protein is a cis-trans prolyl isomerase that binds Observed MW the immunosuppressants FK506 and rapamycin, as well as histone deacetylases, the 30kDa transcription factor YY1, casein kinase II, and nucleolin. It has a higher affinity for rapamycin than for FK506 and thus may be an important target molecule for Calculated MW immunosuppression by rapamycin. 25kDa Category Primary antibody Applications WB, IF, IP Cross-Reactivity Human Recommended Dilutions Immunogen Information WB 1:500 - 1:2000 Gene ID Swiss Prot 2287 Q00688 IF 1:50 - 1:100 Immunogen 1:50 - 1:100 IP Recombinant fusion protein containing a sequence corresponding to amino acids 1-224 of human FKBP3 (NP_002004.1). Synonyms FKBP3;FKBP-25;FKBP-3;FKBP25;PPIase Contact Product Information www.abclonal.com Source Isotype Purification Rabbit IgG Affinity purification Storage Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.02% sodium azide,50% glycerol,pH7.3. Validation Data Western blot analysis of extracts of various cell lines, using FKBP3 antibody (A6907) at 1:1000 dilution. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL Enhanced Kit (RM00021). Exposure time: 30s. Immunoprecipitation analysis of 200ug extracts of MCF-7 cells, using 3 ug FKBP3 antibody (A6907).
    [Show full text]
  • Protein Symbol Protein Name Rank Metric Score 4F2 4F2 Cell-Surface
    Supplementary Table 2 Supplementary Table 2. Ranked list of proteins present in anti-Sema4D treated macrophage conditioned media obtained in the GSEA analysis of the proteomic data. Proteins are listed according to their rank metric score, which is the score used to position the gene in the ranked list of genes of the GSEA. Values are obtained from comparing Sema4D treated RAW conditioned media versus REST, which includes untreated, IgG treated and anti-Sema4D added RAW conditioned media. GSEA analysis was performed under standard conditions in November 2015. Protein Rank metric symbol Protein name score 4F2 4F2 cell-surface antigen heavy chain 2.5000 PLOD3 Procollagen-lysine,2-oxoglutarate 5-dioxygenase 3 1.4815 ELOB Transcription elongation factor B polypeptide 2 1.4350 ARPC5 Actin-related protein 2/3 complex subunit 5 1.2603 OSTF1 teoclast-stimulating factor 1 1.2500 RL5 60S ribomal protein L5 1.2135 SYK Lysine--tRNA ligase 1.2135 RL10A 60S ribomal protein L10a 1.2135 TXNL1 Thioredoxin-like protein 1 1.1716 LIS1 Platelet-activating factor acetylhydrolase IB subunit alpha 1.1067 A4 Amyloid beta A4 protein 1.0911 H2B1M Histone H2B type 1-M 1.0514 UB2V2 Ubiquitin-conjugating enzyme E2 variant 2 1.0381 PDCD5 Programmed cell death protein 5 1.0373 UCHL3 Ubiquitin carboxyl-terminal hydrolase isozyme L3 1.0061 PLEC Plectin 1.0061 ITPA Inine triphphate pyrophphatase 0.9524 IF5A1 Eukaryotic translation initiation factor 5A-1 0.9314 ARP2 Actin-related protein 2 0.8618 HNRPL Heterogeneous nuclear ribonucleoprotein L 0.8576 DNJA3 DnaJ homolog subfamily
    [Show full text]
  • Identification, Quantification and Investigation of Anti-Inflammatory Effects of Echinacea Purpurea Constituents
    KANDHI, VAMSIKRISHNA, Ph.D. Identification, Quantification and Investigation of Anti-inflammatory Effects of Echinacea purpurea Constituents. (2012) Directed by Dr. Nadja B. Cech. 263 pp. Echinacea is among the top selling botanical medicines in the United States. Clinical trials have yielded contradictory reports on the efficacy of Echinacea preparations for treatment of colds, influenza, and inflammation, perhaps due to inconsistency in quality control and lack of information as to which constituents are responsible for their activity. Our overall goal with this research was to develop and apply mass spectrometry techniques to better understand the relationship between chemical composition of Echinacea extracts and their biological activity. This objective was achieved through three separate aims. (1) To determine whether in vitro anti-inflammatory activity of Echinacea purpurea extracts could be correlated with the presence of the alkylamides or caffeic acid derivatives, which have previously been reported to be bioactive. The ability of these extracts to inhibit production of TNF-alpha and PGE2 from influenza A-infected RAW 264.7 macrophage-like cells was assessed. Chemical analysis of the extracts revealed variations in levels of alkylamides, caftaric acid, and cichoric acid. The biological activity of these extracts, however, did not correlate with concentrations of any of these compounds. (2) To investigate the effects of Echinacea purpurea preparations and their constituents on protein expression by immune cells in vitro. Stable isotope labeling with amino acid in cell culture (SILAC), a quantitative proteomics approach, was employed to measure changes in protein expression by activated RAW 264.7 macrophage- type cells exposed to an Echinacea purpurea extract and several of its constituents.
    [Show full text]
  • Inhibition of the FKBP Family of Peptidyl Prolyl Isomerases Induces Abortive Translocation and Degradation of the Cellular Prion Protein
    Inhibition of the FKBP Family of Peptidyl Prolyl Isomerases Induces Abortive Translocation and Degradation of the Cellular Prion Protein by Maxime Sawicki A thesis submitted in conformity with the requirements for the degree of Master of Science Department of Biochemistry University of Toronto © Copyright by Maxime Sawicki 2015 Inhibition of the FKBP Family of Peptidyl Prolyl Isomerases Induces Abortive Translocation and Degradation of the Cellular Prion Protein Maxime Sawicki Master of Science Department of Biochemistry University of Toronto 2015 Abstract Prion disorders are a class of neurodegenerative diseases that feature a structural change of the prion protein from its cellular form (PrPC) into its scrapie form (PrPSc). As these disorders are currently incurable, there is a crucial need for novel therapeutic agents. Here, the FDA-approved immunosuppressive drug FK506 was shown to cause an attenuation in the endoplasmic reticulum (ER) translocation of PrPC by exacerbating an intrinsic inefficiency of PrP’s ER-targeting signal sequence, effectively causing the proteasomal degradation of PrPC. Furthermore, the depletion of FKBP10 also caused the degradation of PrPC but at a later stage following translocation into the ER. Additionally, novel FK506 analogues with reduced immunosuppressive properties were shown to be as efficacious as FK506 in downregulating PrPC. Finally, both FK506 treatment and FKBP10 depletion were shown to reduce the levels of PrPSc in chronically infected cell models. These findings offer a new insight into the development of treatments against prion disorders. ii Acknowledgments The completion of the present thesis would not have been possible without the help and support of a number of people. First and foremost, I would like to thank my supervisor, Dr David Williams, for his constant guidance and expertise that allowed me to successfully complete my degree, as well as my committee members, Dr John Glover and Dr Gerold Schmitt-Ulms, for their invaluable advice and suggestions over the course of this project.
    [Show full text]
  • Downloaded and the Kinasefrom Domainthe Pubmed of TOR Server As Input at the Templates
    biomolecules Review Peptidylprolyl Isomerases as In Vivo Carriers for Drugs That Target Various Intracellular Entities Andrzej Galat Service d’Ingénierie Moléculaire des Protéines (SIMOPRO), CEA, Université Paris-Saclay, F-91191 Gif/Yvette, France; [email protected]; Fax: +33-169089137 Academic Editor: John A. Carver Received: 24 August 2017; Accepted: 26 September 2017; Published: 29 September 2017 Abstract: Analyses of sequences and structures of the cyclosporine A (CsA)-binding proteins (cyclophilins) and the immunosuppressive macrolide FK506-binding proteins (FKBPs) have revealed that they exhibit peculiar spatial distributions of charges, their overall hydrophobicity indexes vary within a considerable level whereas their points isoelectric (pIs) are contained from 4 to 11. These two families of peptidylprolyl cis/trans isomerases (PPIases) have several distinct functional attributes such as: (1) high affinity binding to some pharmacologically-useful hydrophobic macrocyclic drugs; (2) diversified binding epitopes to proteins that may induce transient manifolds with altered flexibility and functional fitness; and (3) electrostatic interactions between positively charged segments of PPIases and negatively charged intracellular entities that support their spatial integration. These three attributes enhance binding of PPIase/pharmacophore complexes to diverse intracellular entities, some of which perturb signalization pathways causing immunosuppression and other system-altering phenomena in humans. Keywords: PPIase; FKBP; cyclophilin; rapamycin; FK506 1. Introduction About forty years ago, two different types of macrocyclic molecules were isolated and shown to possess immunosuppressive activities, such as the cyclic peptide containing non-standard amino acid residues (AAs) cyclosporine A (CsA) and its homologues [1], and two polyketides having L-pipecolic acid ring, namely immunosuppressive macrolide FK506 [2] and rapamycin [3,4].
    [Show full text]
  • The Human Fk506binding Proteins: Characterization of Human FKBP19
    The human FK506-binding proteins: characterization of human FKBP19 Article (Accepted Version) Rulten, Stuart L, Kinloch, Ross A, Tateossian, Hilda, Robinson, Colin, Gettins, Lucy and Kay, John E (2006) The human FK506-binding proteins: characterization of human FKBP19. Mammalian Genome, 17 (4). pp. 322-331. ISSN 0938-8990 This version is available from Sussex Research Online: http://sro.sussex.ac.uk/id/eprint/22944/ This document is made available in accordance with publisher policies and may differ from the published version or from the version of record. If you wish to cite this item you are advised to consult the publisher’s version. Please see the URL above for details on accessing the published version. Copyright and reuse: Sussex Research Online is a digital repository of the research output of the University. Copyright and all moral rights to the version of the paper presented here belong to the individual author(s) and/or other copyright owners. To the extent reasonable and practicable, the material made available in SRO has been checked for eligibility before being made available. Copies of full text items generally can be reproduced, displayed or performed and given to third parties in any format or medium for personal research or study, educational, or not-for-profit purposes without prior permission or charge, provided that the authors, title and full bibliographic details are credited, a hyperlink and/or URL is given for the original metadata page and the content is not changed in any way. http://sro.sussex.ac.uk The Human FK506-Binding Proteins: Characterisation of Human FKBP19 Stuart L Rulten1*, Ross A Kinloch2, Hilda Tateossian3, Colin Robinson2, Lucy Gettins2 and John E Kay4.
    [Show full text]